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Overlapping of mononuclear cells derived from bone marrow in rats' intervertebral discs: an in vitro study

Sobreposição de células mononucleares provenientes da medula óssea em disco intervertebral de ratos: estudo in vitro

Erica Batista Fontes Andréa Pacheco Batista Borges Ney Luis Pippi Maurício Rosa Arícia Sprada Monique Togni About the authors

In this study, bone marrow mononuclear cells (BM-MNC) derived from rats were used in order to promote intervertebral disc regeneration. These cells were isolated after centrifugation in a Ficoll-Paque™ PLUS density gradient and then placed in plastic dishes to proliferate during a period of 14 days. The BM-MNCs were previously labeled with the fluorescent membrane marker Chloromethyl-benzamidodialkylcarbocyanine (CM-DIL), and thereafter were implanted in rats' intervertebral discs explants as an in vitro experimental model. Daily analyses of the cells under a fluorescence microscope revealed morphological changes, which assumed a thin and elongated shape similar to cells that originally form the annulus fibroses. Histopathological analysis demonstrated the presence of mononuclear cells interspersed within collagen fibers. The presence of viable cells, in which were found morphological changes and their disposal in the same pattern of the layers that originate the annulus fibrosus, is an indicator that they engrafted and proliferated on the intervertebral disc. Therefore, morphological changes presented by these cells indicate that they presented mesenchymal stem-like cell characteristics.

mesenchymal stem cells; intervertebral disc; in vitro; rat

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